CN103884800A - Quality detection method of blood-replenishing Angelica sinensis oral liquid - Google Patents
Quality detection method of blood-replenishing Angelica sinensis oral liquid Download PDFInfo
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- CN103884800A CN103884800A CN201410135112.8A CN201410135112A CN103884800A CN 103884800 A CN103884800 A CN 103884800A CN 201410135112 A CN201410135112 A CN 201410135112A CN 103884800 A CN103884800 A CN 103884800A
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Abstract
The invention discloses a quality detection method of blood-replenishing Angelica sinensis oral liquid. The quality detection method comprises the following steps: chromatographic condition and system suitability testing, wherein a chromatographic column is C18, isopropanol-water-glacial acetic acid is used as a mobile phase, the detection wave length is 230nm, the number of theoretical plates is not lower than 2000 according to the paeoniflorin peak, and the flow velocity is 1.5ml/min; preparation of a reference solution, namely precisely weighing a proper amount of paeoniflorin reference substance, and adding methyl alcohol to prepare the solution with the concentration of 40 microgram per 1ml; preparation of a test solution, namely precisely measuring 10ml of Angelica sinensis oral liquid, placing the measured liquid into a 25ml volumetric flask, adding methyl alcohol to obtain constant volume, plugging in a sealing manner, shaking up uniformly, weighting, carrying out ultrasonic treatment for 30min, chilling, weighing again, supplementing the lost weight by methyl alcohol, shaking up, centrifuging at low speed for 10min, and filtering liquid supernatant by a 0.45micron millipore filter membrane, thereby obtaining subsequent filtrate; and precisely drawing 10 micro liter of reference solution and 10 micro liter of test solution respectively, injecting into a liquid chromatograph and determining. The method is simple, convenient and accurate, and is good in repeatability and high in specificity.
Description
Technical field
The invention belongs to technical field of traditional Chinese medicines, particularly the quality determining method of Chinese angelica oral liquid for benefiting blood.
Background technology
Chinese angelica oral liquid for benefiting blood is the change formulation product carrying out on the basis of Radix Angelicae Sinensis benefit blood cream, is a kind of Chinese medicine pure preparation.The oral liquid being made into, had both retained former formulation and had absorbed soon, the feature rapidly of proving effective; There is mouthfeel good simultaneously, easy for patients to accept; In finished product, add suitable antiseptic, and through sterilization treatment, pack steady quality; Make the oral liquid of single dose, taking convenience, dosage is accurate, and the plurality of advantages such as be easy to carry simultaneously.Chinese angelica oral liquid for benefiting blood by Radix Angelicae Sinensis, the root of herbaceous peony (wine system), Ligusticum wallichii (wine system), Radix Codonopsis, the Radix Astragali (honey is processed), donkey-hide gelatin, Poria cocos, Radix Glycyrrhizae through water extraction get concentrated after, add sucrose, Sodium Benzoate preparation, filling forming.There is effect of nourishing qi and blood, for anaemia, dizziness, palpitation and amnesia, irregular menstruation, postpartum the deficiency of blood, weak.Assay in this product proper mass standard, to detect Radix Angelicae Sinensis, the contained forulic acid of Ligusticum chuanxiong Hort in prescription, but find in actual production, storage, survey index using forulic acid as containing, production quality control is had to following some weak point: 1, forulic acid is extremely unstable in this product, easily decompose.The long-term reserved sample observing discovery of this product, product content declines fast, contrasts Paeoniflorin accelerated test and long-term stable experiment simultaneously, and the every monthly average degradation rate of forulic acid reaches 10%, and Paeoniflorin is more stable, degradation rate is less than 5%.Meanwhile, a large amount of documents and materials are also reported in the recent period, and the forulic acid solution of water or methyl alcohol (ethanol) preparation is unstable, is subject to the impact of light, heat, pH, metallic ion etc., very easily decompose, thereby reduce check accuracy test; 2, survey index using forulic acid as containing, in finished product, the rate of transform of forulic acid is very low, and finished product content standard is only for 0.3mg/ props up.Because content is very low, thereby be unfavorable for controlling Chinese angelica oral liquid for benefiting blood quality.
Summary of the invention
A kind of easy, accurate, the favorable reproducibility that the object of the invention is to overcome above-mentioned shortcoming and provide, the quality determining method of the strong Chinese angelica oral liquid for benefiting blood of specificity.
The quality determining method of a kind of Chinese angelica oral liquid for benefiting blood of the present invention, comprises the steps:
(1) chromatographic condition and system suitability test: chromatographic column is C18(250 mm × 4.6 mm, and 5 μ m); Isopropanol-water-glacial acetic acid (4.8:95:0.2) is mobile phase; Detection wavelength is 230nm.Theoretical cam curve is calculated and should be not less than 2000 by Paeoniflorin peak; Flow velocity: 1.5ml/min;
(2) preparation of reference substance solution: get Paeoniflorin reference substance appropriate, accurately weighed, add methyl alcohol and make the solution of every 1ml containing 40 μ g, to obtain final product;
(3) preparation of need testing solution: precision measures this product 10ml, puts in 25ml volumetric flask, adds methanol constant volume, close plug, shakes up, weighed weight, ultrasonic 30 minutes, let cool, more weighed weight, supply the weight of less loss with methyl alcohol, shake up, low-speed centrifugal 10min, gets supernatant and filters with 0.45 μ m miillpore filter, get subsequent filtrate, to obtain final product;
(4) determination method: accurate reference substance solution and the each 10 μ l of need testing solution of drawing respectively, injection liquid chromatography, measures.
The quality determining method of above-mentioned a kind of Chinese angelica oral liquid for benefiting blood, wherein: every of this product, in Paeoniflorin, must not be less than 0.9mg.
The present invention compared with prior art, there is obvious beneficial effect, from above technical scheme: the present invention is using Paeoniflorin as assay index, in its finished product, the Paeoniflorin rate of transform is higher, reach 56% left and right, select content standard must not be less than 0.9mg/ and prop up, higher than ferulaic acid content limit standard; And Paeoniflorin has stronger pharmacologically active and good physical and chemical stability, can effectively control product quality.In order to ensure detection method authenticity, reflection product quality, effective control for product quality.
Embodiment
Further illustrate below the beneficial effect of the inventive method by specific experiment example.
experimental example:
1 instrument and reagent
High performance liquid chromatograph (pump: LC-20AD, detecting device: SPD-M20A); Workstation: LC Solution.Chinese angelica oral liquid for benefiting blood (Guiyang Xintian Pharmaceutical Industry Co., Ltd.), and Paeoniflorin reference substance (for assay, Nat'l Pharmaceutical & Biological Products Control Institute, lot number: 110736-200934, content is in 95.7%); Water is redistilled water, and glacial acetic acid, phosphoric acid are that analysis is pure, and methyl alcohol, acetonitrile, isopropyl alcohol are chromatographically pure.
2 methods and result
2.1 chromatographic condition chromatographic columns are C18(250 mm × 4.6 mm, 5 μ m), taking isopropanol-water-glacial acetic acid (4.8:95:0.2) as mobile phase; Detection wavelength is 230nm.Theoretical cam curve is calculated and should be not less than 2000 by Paeoniflorin peak.Flow velocity: 1.5ml/min.
It is appropriate that Paeoniflorin reference substance is got in the preparation of 2.2 reference substance solution, accurately weighed, adds methyl alcohol and make the solution of every 1 ml containing 40 μ g.
The preparation precision of 2.3 need testing solutions measures this product 10ml, puts in 25ml volumetric flask, adds methanol constant volume, close plug, shakes up, weighed weight, ultrasonic 30 minutes, let cool, more weighed weight, supply the weight of less loss with methyl alcohol, shake up, low-speed centrifugal 10min, gets supernatant and filters with 0.45 μ m miillpore filter, get subsequent filtrate, to obtain final product.
The preparation of 2.4 negative control solutions is got other medicinal materials of removing white Peony Root and is prepared blank sample in prescription ratio according to production technology preparation method, makes negative control product solution by " 2.3 " below method.
Accurate need testing solution, reference substance solution, the negative control solution drawn of 2.5 system suitabilities, by 2.1 lower chromatographic condition sample introduction 10 μ l, record chromatogram, there is chromatographic peak with Paeoniflorin reference substance in corresponding position in result test sample, and degree of separation is good; Negative control occurs without chromatographic peak in this position.
2.6 linear relationships are investigated precision and are taken Paeoniflorin reference substance 16.56mg(lot number 110736-200934, and content is in 95.7%), be placed in 100ml volumetric flask, add methyl alcohol and dissolve and be diluted to scale, obtain the mother liquor of every 1ml containing 158.48 μ g.Precision measures this mother liquor 5ml, puts in 20ml volumetric flask, adds methyl alcohol and is diluted to scale, shakes up, and obtains the reference substance solution of every 1ml containing 39.62 μ g.According to above-mentioned chromatographic condition with reference substance solution 3 μ l, 5 μ l, 10 μ l, 15 μ l, 20 μ l, 25 μ l sample introductions, measure peak area, taking peak area as ordinate, Paeoniflorin sample size is horizontal ordinate drawing standard curve, and fitting a straight line equation is: Y=9114.47X-976.8; The straight-line equation that initial point is crossed in matching is y=9100.6x R2=0.9995295 response relative standard deviation (RFRSD)=1.62459, and within the scope of definite 11.886 μ g～99.049 μ g, the sample size of Paeoniflorin reference substance and peak area value are good linear relationship thus.
The accurate Paeoniflorin reference substance solution of drawing of 2.7 precision tests, repeats sample introduction 6 times by above-mentioned chromatographic condition, records chromatogram, result Paeoniflorin reference substance peak area RSD=1.08%.
Same batch sample is got in 2.8 reappearance tests, makes 6 parts of need testing solutions by the preparation method under " 2.3 " item, and sample introduction, records chromatogram respectively, calculates result RSD=1.28%.
2.9 stability degree tests are placed need testing solution after 0,2,4,6,8,12 hour, the accurate each 10 μ l of need testing solution that place different time points that draw respectively, injection liquid chromatography, record chromatogram, in result need testing solution Paeoniflorin in 12 h peak area without significant change, result RSD=1.72%.
2.10 average recoveries tests are accurate draws (111001) 9 parts, sample having measured content, every part of 5ml(content: 1.2mg/ props up), wherein 1,2, No. 3 three parts respectively add reference substance 495.89 μ g, 4,5, No. 6 three parts respectively add reference substance 644.20 μ g, 7,8, No. 9 three parts respectively add reference substance 726.98 μ g, prepare solution by the preparation method under " 2.3 " item, sample introduction, records chromatogram, calculate recovery rate respectively.The results are shown in Table 1.
2.11 sample sizes are measured and are got Chinese angelica oral liquid for benefiting blood, make need testing solution by the preparation method under " 2.3 " item, measure by this paper chromatographic condition, record chromatogram, calculate content.The results are shown in Table 2.
Definite statistics sample of 2.12 content limits and the medicinal material content using, calculating mean transferred rate is 55.58%
According to the content limit of " Chinese Pharmacopoeia " 2010 editions middle white Peony Root Paeoniflorins, in conjunction with the Paeoniflorin rate of transform in white Peony Root recipe quantity in this product and upper table, be calculated as follows the content limit that draws this product:
Be that 0.91mg/ props up according to above-mentioned computing formula result of calculation, therefore specify that every of this product contains the root of herbaceous peony with Paeoniflorin (C
23h
28o
11) meter, must not be less than 0.9mg/ and prop up.
3 discuss
In we, when being classified as main ingredient, forulic acid is main effective constituent in Radix Angelicae Sinensis, but forulic acid is unstable, easily decomposes, therefore select in the root of herbaceous peony Paeoniflorin as the composition of this law control.
Measure with reference to paeoniflorin content in white Peony Root of " Chinese Pharmacopoeia " version in 2010, wavelength is 230nm; Paeoniflorin reference substance UV scanning maximum absorption wavelength is 231.40nm.Therefore drafting and detecting wavelength is 230nm.
In need testing solution preparation, to refluxing extraction and ultrasonic comparing, result refluxing extraction and ultrasonic extraction do not have significant difference, select ultrasonic extraction at this.Ultrasonic extraction times 10～40 min is compared, and result shows to extract after 30 min, and in sample, Paeoniflorin extracts completely substantially, therefore at these ultrasonic extraction 30 min.
Because this product is without alcohol precipitation, in the time that filtering with 0.45 μ m miillpore filter, the sample of ultrasonic processing finds more sad filter, therefore low-speed centrifugal 10min after ultrasonic processing removes precipitation, be convenient to filter with miillpore filter.
embodiment:
A quality determining method for Chinese angelica oral liquid for benefiting blood, comprises the steps:
(1) chromatographic condition and system suitability test: chromatographic column is C18(250 mm × 4.6 mm, and 5 μ m); Isopropanol-water-glacial acetic acid (4.8:95:0.2) is mobile phase; Detection wavelength is 230nm.Theoretical cam curve is calculated and should be not less than 2000 by Paeoniflorin peak; Flow velocity: 1.5ml/min;
(2) preparation of reference substance solution: get Paeoniflorin reference substance appropriate, accurately weighed, add methyl alcohol and make the solution of every 1ml containing 40 μ g, to obtain final product;
(3) preparation of need testing solution: precision measures this product 10ml, puts in 25ml volumetric flask, adds methanol constant volume, close plug, shakes up, weighed weight, ultrasonic 30 minutes, let cool, more weighed weight, supply the weight of less loss with methyl alcohol, shake up, low-speed centrifugal 10min, gets supernatant and filters with 0.45 μ m miillpore filter, get subsequent filtrate, to obtain final product;
(4) determination method: accurate reference substance solution and the each 10 μ l of need testing solution of drawing respectively, injection liquid chromatography, measures, and every of this product contains the root of herbaceous peony in Paeoniflorin, must not be less than 0.9mg.
Claims (2)
1. a quality determining method for Chinese angelica oral liquid for benefiting blood, comprises the steps:
(1) chromatographic condition and system suitability test: chromatographic column is C18; Volume ratio=the 4.8:95:0.2 of isopropanol-water-glacial acetic acid is mobile phase; Detection wavelength is 230nm; Theoretical cam curve is calculated and should be not less than 2000 by Paeoniflorin peak; Flow velocity: 1.5ml/min;
(2) preparation of reference substance solution: get Paeoniflorin reference substance appropriate, accurately weighed, add methyl alcohol and make the solution of every 1ml containing 40 μ g, to obtain final product;
(3) preparation of need testing solution: precision measures this product 10ml, puts in 25ml volumetric flask, adds methanol constant volume, close plug, shakes up, weighed weight, ultrasonic 30 minutes, let cool, more weighed weight, supply the weight of less loss with methyl alcohol, shake up, low-speed centrifugal 10min, gets supernatant and filters with 0.45 μ m miillpore filter, get subsequent filtrate, to obtain final product;
(4) determination method: accurate reference substance solution and the each 10 μ l of need testing solution of drawing respectively, injection liquid chromatography, measures.
2. the quality determining method of a kind of Chinese angelica oral liquid for benefiting blood as claimed in claim 1, wherein: every of this product, in Paeoniflorin, must not be less than 0.9mg.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101703611A (en) * | 2009-11-30 | 2010-05-12 | 贵阳新天药业股份有限公司 | Quality detection method of Chinese angelica oral liquid for benefiting blood |
| CN101926889A (en) * | 2010-08-09 | 2010-12-29 | 四川美大康药业股份有限公司 | Method for detecting white paeony root-medlar particles |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101703611A (en) * | 2009-11-30 | 2010-05-12 | 贵阳新天药业股份有限公司 | Quality detection method of Chinese angelica oral liquid for benefiting blood |
| CN101926889A (en) * | 2010-08-09 | 2010-12-29 | 四川美大康药业股份有限公司 | Method for detecting white paeony root-medlar particles |
Non-Patent Citations (4)
| Title |
|---|
| HUA-BIN LI 等: "Separation methods used for Scutellaria baicalensis active components", 《JOURNAL OF CHROMATOGRAPHY B》, vol. 812, no. 12, 5 December 2004 (2004-12-05), pages 277 - 290, XP004648225, DOI: doi:10.1016/j.jchromb.2004.06.045 * |
| 周祖坤 等: "高效液相色谱法测定当归芍药颗粒中芍药苷的含量", 《药物分析杂志》, vol. 26, no. 7, 31 July 2006 (2006-07-31), pages 1001 - 1003 * |
| 胡丹 等: "高效液相色谱法测定归芍调经胶囊中芍药苷的含量", 《安徽医药》, vol. 15, no. 1, 31 January 2011 (2011-01-31), pages 42 - 43 * |
| 陈亚林 等: "高效液相色谱法测定妇炎康丸中芍药苷的含量", 《中国医院药学杂志》, vol. 32, no. 11, 15 June 2012 (2012-06-15), pages 899 - 900 * |
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